Cryopreservation method of cranoglanis bouderius seminal fluid and diluent thereof
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A technology of ultra-low temperature preservation and diluent, which is applied to the preservation, application, and animal husbandry of human or animal bodies, and achieves remarkable effects, improved semen protection, and considerable ecological, economic, and social benefits
Active Publication Date: 2017-08-18
SOUTH CHINA AGRI UNIV +2
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Problems solved by technology
[0004] The technical problem to be solved by the present invention is to overcome the deficiencies in the existing semen preservation technology of S. longiscus, and provide a semen dilution and ultra-low temperature preservation method for germplasm research, genetic management, semen cryopreservation and germplasm protection. provide guarantees, and provide important scientific guidance for solving the technical problems of artificial propagation of the long-breasted catfish, protecting the resources of the long-breasted catfish, and realizing the healthy and sustainable development of my country's famous and high-quality fish farming industry.
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Embodiment 1
[0052] Example 1 The semen preservation diluent of the long-breasted catfish
[0053] 1. Dilution of sperm preservation solution Formula A: Prepare solution A and solution B separately. According to our exploration and experiments, the best formula of solution A is: glucose 1.80g, NaCl 15.2g, KCl 1.68g, CaCl 2 0.28g, MgSO 4 ·7H 2 O 0.472g, finally add 100ml of distilled water to dissolve;
[0054] The formula of liquid B is NaHCO 3 0.65g, Na 2 HPO 4 ·H 2 O 0.35g, KH 2 PO 4 0.12g, C 6 h 12 o 6 ·H 2 O 1.88g, finally add 100ml of distilled water to dissolve.
[0055] Take 10 mL each of Solution A and Solution B, mix them, add distilled water to make up to 100 mL, and add dimethyl sulfoxide with a final volume concentration of 5%, 10% or 15% to prepare sperm preservation dilution A.
[0056] 2. Sperm Preservation Diluent B Formula: Glucose 1.80g, Fructose 1.2g, KCl 0.08g, CaCl 2 0.02g, NaHCO 3 0.21g, distilled water 100mL, and add dimethyl sulfoxide with a final ...
Embodiment 2
[0059] Example 2 The cryopreservation method of the semen of the long-breasted catfish
[0060] The invention provides a method for cryopreservation of semen of C. elongatus. The main steps are as follows: selection of fish species, preparation of diluent for sperm preservation and antifreeze, sperm collection and quality observation, cryopreservation of semen, thawing and recovery, and observation of sperm viability. The specific operation method is as follows:
[0061] 1. Fish species selection
[0062] The collection of fish fingerlings in the natural environment is an important source of broodstock for the long-breasted catfish. Through long-term field research, we have introduced and collected more than 300 broodstock of the long-breasted catfish. More than 100 individuals weighing more than 100 catties have been intensively cultivated in the scientific research and demonstration base of Qingyuan Xingyu Aquatic Technology Co., Ltd. When collecting semen, choose male broo...
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Abstract
The invention discloses a preserved diluent of cranoglanis bouderius seminal fluid and a cryopreservation method of the cranoglanis bouderius seminal fluid. The preserved diluent is prepared from sugar, sodium salt, potassium salt, calcium salt, tyrosine, glycine, distilled water and an anti-freezing agent-dimethyl sulfoxide. The cryopreservation method comprises the following steps: selecting good male parent fish meeting the conditions to carry out seminal fluid collection, preserving in the preserved diluent in which neomycin sulfate and streptomycin are added, adopting a cooling rate which is -10.5 DEG C/30 seconds, and putting the seminal fluid in liquid nitrogen for cryopreservation after reaching -150 DEG C; when unfreezing and recovering are needed, taking out the cyropreserved cranoglanis bouderius seminal fluid, adding a PBS (Phosphate Buffer Solution) of which the volume percentage is 0.65 to 0.7 percent as an unfreezing solution, and putting the cranoglanis bouderius seminal fluid in a water bath of which the temperature is 32 to 40 DEG C for rapid unfreezing and recovering. According to the technology disclosed by the invention, the durability of preservation time of the cranoglanis bouderius seminal fluid can be ensured, the protection effect of the seminal fluid is good, and sperms after preservation are high in survival rate and good in vitality.
Description
technical field [0001] The invention belongs to the technical field of aquaculture. More specifically, it relates to a method for cryopreservation of the semen of the rare and endangered fish, the elongated catfish, and its dilution. Background technique [0002] The long-breasted catfish is a fish belonging to the family Siluriformes and the long-breasted catfish. Body long, laterally flattened, with dorsal fin starting at the highest point of the body. The head is flat, slightly triangular in shape, and the back bone is rough and exposed. Snout prominent, blunt. Mouth proximal, arc-shaped, upper jaw slightly protruding. It is a bottom fish of subtropical foothill rivers and streams, and likes clear and flowing water environment. Gluteus longus has no intermuscular thorns, delicious meat and high nutritional value, so the market price is relatively high, and the supply is often in short supply. Due to the overfishing of the wild echinacea, its wild resources and seedl...
Claims
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